Horses may be carriers of important resistant bacteria like methicillin-resistant staphylococci. Such bacteria can potentially threaten both equine and public health, but little is known about predisposing factors like antimicrobial usage patterns in equines. Objectives of this study were to investigate the antimicrobial usage practices by Danish equine practitioners as well as factors impacting usage. A total of 103 equine practitioners filled in an online questionnaire. When asked to explain their typical treatment of six clinical case scenarios, only 1% and 7% of respondents prescribed systemic antimicrobials for a cough and pastern dermatitis, respectively. More frequent usage was reported for diarrhoea (43%), extraction of a cracked tooth (44%), strangles (56%), and superficial wound near a joint (72%). Among the antibiotics indicated for treatment, enrofloxacin was the only critically important antimicrobial agent reported by two respondents. Thirty-eight (36%) respondents worked in practices with antimicrobial protocols. When asked to prioritize the most important factor influencing prescribing habits, bacterial culture (47%) and antimicrobial protocols (45%) were selected far more often than the owner´s economy (5%) and expectations (4%). Veterinarians reported limitations such as the availability of only one oral antibiotic (sulphadiazine/trimethoprim), and a need for clearer treatment guidelines. In conclusion, the study highlighted important aspects regarding antimicrobial usage among equine practitioners. Antimicrobial protocols and pre- and post-graduate education on prudent antimicrobial usage are recommended.